羟基磷灰石表面台阶结构对未成熟树突状细胞mRNA表达谱的影响  

The Effects of Stepped Structures of Hydroxyapatite Surface on the mRNA Expression Profile of Immature Dendritic Cells

作　　者：王芳 陈丽丹 王赟 胡祖权 曾柱 陈晋 WANG Fang;CHEN Lidan;WANG Yun;HU Zuquan;ZENG Zhu;CHEN Jin(Key Laboratory of Infectious Immune and Antibody Engineering in University of Guizhou Province,School of Basic Medical Sciences,Guizhou Medical University,Guiyang,550025;Immune Cells and Antibody Engineering Research Center in University of Guizhou Province,School of Biology and Engineering(School of Modern Industry for Health and Medicine),Guizhou Medical University,Guiyang,550025)

机构地区：[1]贵州医科大学基础医学院,贵州省高等学校感染免疫与抗体工程重点实验室,贵阳550025 [2]贵州医科大学生物与工程学院(健康医药现代产业学院),贵州省高等学校免疫细胞与抗体工程研究中心,贵阳550025

出　　处：《基因组学与应用生物学》2024年第9期1632-1645,共14页Genomics and Applied Biology

基　　金：国家自然科学基金项目(32160226,12132006);贵州省科技计划项目黔科合平台人才[2021]5637号项目共同资助。

摘　　要：羟基磷灰石(hydroxyapatite,HA)在骨修复和材料界面修饰中具有重要价值,但其在植入体内后可能会引起慢性炎症,而树突状细胞(dendritic cells,DCs)是调控慢性炎症反应的关键细胞,其表型和功能受材料表面结构影响。因此,本研究在构建HA表面台阶结构的基础上,分析其对未成熟树突状细胞(immature dendritic cells,imDCs)的细胞骨架结构、黏附形态以及mRNA表达谱的影响,以期为优化HA材料的表面结构设计提供一定的理论依据。利用取向连接生长原理制备出具有台阶结构的HA片材,并以此构建细胞培养模型。利用免疫荧光染色分析imDCs细胞骨架和黏附形态的变化,并采用转录组测序技术分析imDCs的mRNA表达谱的变化。结果显示,HA表面台阶结构的暴露会增加imDCs的铺展面积,并影响细胞黏附形态。转录组测序分析显示mRNA表达谱发生了变化,差异表达基因(differentially expressed genes,DEGs)主要富集到黏着斑信号通路、趋化因子信号通路、白介素-17信号通路等关键信号通路。因此,HA台阶结构可能通过黏着斑信号通路、趋化因子信号通路、白介素-17信号通路调控imDCs的形态特征和免疫学功能,进而影响免疫应答过程。Hydroxyapatite(HA)has significant clinical promise for the repair of bone defects and the modification of implant material interfaces.However,after implantation,there are potential risks of chronic inflammation.Dendritic cells(DCs)are key cells in regulating chronic inflammatory responses,and their phenotype and functions are influenced by the surface structure of materials.Therefore,in order to provide some theoretical guidance for the optimal design of HA surface structures,this study tried to construct stepped structures on HA surfaces and investigate the effects of stepped structures on the cytoskeletal,cellular morphology,and mRNA expression profile of immature dendritic cells(imDCs).HA dishes with stepped structures were prepared utilizing the principle of oriented attachment growth,and subsequently used to construct in vitro cell culture models.The imDCs were observed by immunofluorescence staining to characterize their cytoskeleton structures and cellular morphology.Besides,transcriptome sequencing was used to examine the changes in the mRNA expression profile of imDCs.The results indicated that the stepped structures exposed on HA surfaces would increase the spread area of imDCs and affect the cell adhesion morphology.In addition,the results of the transcriptome sequencing revealed significant alterations in the mRNA expression profile,with differentially expressed genes(DEGs)predominantly enriched in the pathways associated with the focal adhesion signaling pathway,the chemokine signaling pathway,and the IL-17 signaling pathway.Thus,the stepped structures could modulate the cellular morphology and the immunological functions of imDCs by modulating the aforementioned signaling pathways,including the focal adhesion pathway,the chemokine signaling pathway,and the IL-17 signaling pathway,thereby affecting the immune response process.

关 键 词：羟基磷灰石 树突状细胞 台阶结构 转录组测序 信号通路 

分 类 号：R318.08[医药卫生—生物医学工程]